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Open AccessArticle

Screening of Differentially Expressed Microsporidia Genes from Nosema ceranae Infected Honey Bees by Suppression Subtractive Hybridization

1
Department of Biotechnology and Animal Science, National Ilan University, No. 1, Sec. 1, Shen Nung Road, Ilan 26047, Taiwan
2
Genomics Research Center, Academia Sinica, No. 128, Academia Road, Sec. 2, Nankang District, Taipei 115, Taiwan
3
Department of Entomology, National Chung Hsing University, No. 145, Xingda Road, Taichung 402, Taiwan
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Insects 2020, 11(3), 199; https://doi.org/10.3390/insects11030199
Received: 21 February 2020 / Revised: 17 March 2020 / Accepted: 18 March 2020 / Published: 22 March 2020
The microsporidium Nosema ceranae is a high prevalent parasite of the European honey bee (Apis mellifera). This parasite is spreading across the world into its novel host. The developmental process, and some mechanisms of N. ceranae-infected honey bees, has been studied thoroughly; however, few studies have been carried out in the mechanism of gene expression in N. ceranae during the infection process. We therefore performed the suppressive subtractive hybridization (SSH) approach to investigate the candidate genes of N. ceranae during its infection process. All 96 clones of infected (forward) and non-infected (reverse) library were dipped onto the membrane for hybridization. A total of 112 differentially expressed sequence tags (ESTs) had been sequenced. For the host responses, 20% of ESTs (13 ESTs, 10 genes, and 1 non-coding RNA) from the forward library and 93.6% of ESTs (44 ESTs, 28 genes) from the reverse library were identified as differentially expressed genes (DEGs) of the hosts. A high percentage of DEGs involved in catalytic activity and metabolic processes revealed that the host gene expression change after N. ceranae infection might lead to an unbalance of physiological mechanism. Among the ESTs from the forward library, 75.4% ESTs (49 ESTs belonged to 24 genes) were identified as N. ceranae genes. Out of 24 N. ceranae genes, nine DEGs were subject to real-time quantitative reverse transcription PCR (real-time qRT-PCR) for validation. The results indicated that these genes were highly expressed during N. ceranae infection. Among nine N. ceranae genes, one N. ceranae gene (AAJ76_1600052943) showed the highest expression level after infection. These identified differentially expressed genes from this SSH could provide information about the pathological effects of N. ceranae. Validation of nine up-regulated N. ceranae genes reveal high potential for the detection of early nosemosis in the field and provide insight for further applications. View Full-Text
Keywords: Microsporidia; Nosema ceranae; honey bee; cDNA subtraction Microsporidia; Nosema ceranae; honey bee; cDNA subtraction
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Chang, Z.-T.; Ko, C.-Y.; Yen, M.-R.; Chen, Y.-W.; Nai, Y.-S. Screening of Differentially Expressed Microsporidia Genes from Nosema ceranae Infected Honey Bees by Suppression Subtractive Hybridization. Insects 2020, 11, 199.

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